RU2399622C2 - Novel porphyrin derivatives, particularly chlorins and/or bacteriochlorins and use thereof in photodynamic therapy - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to porphyrin derivatives of formula

and

, where X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸ denote halogen atoms or hydrogen atoms and R¹, R², R³, R⁴ are selected from OH-groups, amino acids, OR-groups, NHR-groups and/or chlorine atoms, where R is an alkyl containing 1-12 carbon atoms. The invention also relates to a method of obtaining said compounds and medicinal agents prepared from said compounds.

EFFECT: possibility of using compounds and medicinal agents in photodynamic therapy of cancerous diseases.

19 cl, 2 ex

Description

State of the art

Photodynamic therapy (PDT) is a method used to treat certain types of diseases, in particular certain types of cancer. This technique consists in introducing a photosensitizer into the pathological tissues as a label, then causing selective destruction of these tissues, exposing these tissues to light with a specific wavelength. Such monochromatic light is usually obtained using a laser or laser diode.

A photosensitizer is understood to mean any molecule that is capable of storing light energy, which can be activated by the indicated energy, and therefore provide itself for numerous biochemical combinations.

Typically, a photosensitizer is administered to a patient in an amount of between 0.1 and 0.5 micromoles per 1 kg of body weight, then cells of the whole organism catch it. The photosensitizing molecule accumulates mainly in cancer cells, but remains inactive until it is exposed to light with a suitable wavelength. Illumination or irradiation of the tumor with light activates a molecule, which then interacts with oxygen, and short-lived compounds are formed, in particular singlet oxygen. Singlet oxygen is a highly reactive and toxic molecule that destroys the cancer cells in which the photosensitizer has been concentrated.

Some time passes between the introduction of the photosensitizer and its activation by the laser. The laser light used in photodynamic therapy is focused with an optical fiber and exposed to it for only a few minutes. The person performing this procedure holds the optical fiber in close proximity to the tumor in order to deliver the exact amount of light. As a result, photodynamic therapy only minimally destroys healthy cells.

In the initial stage of cancer, the goal of such a technique may be to completely eradicate and cure the cancer, but in the late stage, the task may be to reduce the volume of the tumor in order to alleviate the symptoms. New normal cells are replaced by those that were destroyed by photodynamic therapy, which ensures quick recovery after treatment and excludes, in particular, gross scars that can form as a result of tissue removal by other methods.

Even if patients have already been treated with surgical methods, radiotherapy or chemotherapy methods, this technique can undoubtedly be applied to them.

Examples of diseases that can be treated with photodynamic therapy include, but are not limited to, stomach cancer, intestinal cancer, lung cancer, breast cancer, uterine cancer, esophageal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, gall bladder cancer, tongue cancer, brain cancer, skin cancer, thyroid cancer, prostate cancer, parotid cancer, as well as some viral and / or microbial diseases.

Photodynamic therapy can be used as a way to diagnose certain forms of cancer. From this point of view, it is sufficient that the photosensitizing molecules fluoresce and, therefore, are able to emit light after they have been irradiated.

It is currently known that in photodynamic therapy some derivatives of porphyrins, in particular hematoporphyrin, are used as photosensitizers. This remedy is known under the registered AXCAN PHARMA INC. Photofrin trademark is a purified mixture of hematoporphyrins. Hematoporphyrin is, in turn, a derivative of porcine hemoglobin.

In addition, Photofrin® has so far been the only photosensitizing molecule that has been approved for sale in some countries, particularly France, for the treatment of esophageal cancer.

Despite the advantages demonstrated by Photofrin®, such as solubility in an aqueous medium, good yield in the formation of singlet oxygen and easy synthesis, it still has several disadvantages.

First of all, Photofrin® is activated with light with a wavelength of 630 nm. However, at this wavelength, light penetrates tissue only in the range of 5 to 10 mm, which is a serious obstacle when the tumor is more extensive and lies deeper. Moreover, such a photosensitizer leads to skin photosensitivity within six weeks after application. Finally, the fact that Photofrin® is a mixture of several molecules makes it more difficult to select the appropriate dosage for both the photosensitizer and the light supplied.

Among other test compounds, the so-called second-generation compounds, document WO-98/50386 describes some benzoporphyrin derivatives. Such derivatives demonstrate some advantages over Photofrin® in that they absorb light at a wavelength of 690 nm and therefore can be used in the treatment of larger and deeper tumors. Such derivatives, in addition, have better selectivity for cancer cells.

However, certain problems remain related to selectivity, light absorption and toxicity.

An object of the present invention is to propose new derivatives of porphyrin, such as, in particular, chlorins and / or bacteriochlorins, which have eliminated the aforementioned disadvantages, in particular with respect to preferred accumulation in cancer cells, better stability and lower phototoxicity.

Another objective of the present invention is the proposal of anticancer, and / or antiviral, and / or antimicrobial agents for the purpose of their use in photodynamic therapy.

Other objectives and advantages of the invention will become apparent in the following description, which is given by way of example only and not limited to.

Disclosure of invention

The present invention relates to derivatives of porphyrin, in particular to chlorine, a compound having the following formula:

Where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, are selected from halogen atoms and / or hydrogen atoms;

- R ¹ , R ² , R ³ , R ⁴ , each, identical or different, are selected from the following substituents: OH groups, amino acids, OR groups and NHR groups and / or a chlorine atom, where R is an alkyl group, having from 1 to 12 carbon atoms.

The present invention also relates to porphyrin derivatives, in particular bacteriochlorin, a compound having the following formula:

Where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, are selected from halogen atoms and / or hydrogen atoms;

- R ¹ , R ² , R ³ , R ⁴ , each, identical or different, are selected from OH groups, amino acids, OR groups and NHR groups and / or a chlorine atom, where R is an alkyl group comprising from 1 up to 12 carbon atoms.

One of the advantages of these derivatives is that, as described in the present invention, a chlorosulfonyl group is present in the molecule, which makes it possible to easily and efficiently introduce a wide variety of chemical substituents.

In addition, the present invention relates to anticancer and / or antiviral and / or antimicrobial drugs used to treat humans or animals, containing, as the main active substance, one or more compounds described in the present invention.

Brief Description of the Drawings

The invention will be better understood by reading the following description, accompanied by the accompanying drawings, including:

- Figure 1 shows the absorption spectrum of chlorin, a compound having the formula (I) in an aqueous medium before (solid line) and after (dashed line) irradiation of the sample with the second harmonic of the Nd: YAG laser.

- Figure 2 shows the absorption spectrum of bacteriochlorin, a compound having the formula (II), in an aqueous medium before (solid line) and after (dashed line) irradiation of the sample with the second harmonic of the Nd: YAG laser.

The implementation of the invention

The present invention, firstly, relates to derivatives of porphyrin, in particular to chlorine, a compound having the following formula:

Where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, selected from halogen atoms and / or hydrogen atoms;

- R ¹ , R ² , R ³ , R ⁴ , each, identical or different, are selected from the following substituents: OH groups, amino acids, OR groups and NHR groups and / or chlorine atoms, where R is an alkyl group, having from 1 to 12 carbon atoms.

One of the advantages of these types of derivatives is that they are amphiphilic substances. Amphiphilicity is an essential feature, since such molecules can cross cell membranes and accumulate inside cells.

An amphiphilic compound is understood to mean any molecule that exhibits both hydrophilic and hydrophobic properties.

This type of derivative has an absorption band at 650 nm, as shown in FIG.

The present invention also relates to derivatives satisfying the formula (I), where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, are selected from halogen atoms and / or hydrogen atoms, so that at least one of X radicals in each pair of X ¹ / X ² , X ³ / X ⁴ , X ⁵ / X ⁶ , X ⁷ / X ⁸ is a halogen atom.

The advantage of this particular type of derivatives lies in the presence of halogen atoms in the ortho position of the phenyl substituent, which increases the quantum yield of the formation of the triplet state without significantly reducing its half-life. With a higher quantum yield, the triplet state can transfer its energy more efficiently than oxygen with a correspondingly higher yield in the formation of singlet oxygen.

In addition, the halogen atoms in the ortho positions of the phenyl substituent make the tetrapyrrole macrocycle more stable and, therefore, more effective when used in photodynamic therapy.

In addition, according to the present invention and in accordance with formula (I):

- X ² , X ⁴ , X ⁶ and X ⁸ are either chlorine atoms or fluorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are chlorine atoms.

According to another embodiment of the present invention and in accordance with formula (I):

- X ² , X ⁴ , X ⁶ and X ⁸ are either chlorine atoms or fluorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are OH groups.

The present invention also relates to derivatives corresponding to the formula (I), where:

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms,

- R ¹ , R ² , R ³ , R ⁴ are either chlorine atoms or OH groups.

The present invention also relates to porphyrin derivatives, in particular bacteriochlorin, a compound having the following formula:

Where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, are selected from halogen atoms and / or hydrogen atoms;

- R ¹ , R ² , R ³ , R ⁴ , each, identical or different, are selected from the following substituents: OH groups, amino acids, OR groups and NHR groups and / or a chlorine atom, where R is an alkyl group, having from 1 to 12 carbon atoms.

The advantage of this type of derivatives and those described above for chlorins is that they are also amphiphilic.

Another advantage of these derivatives, in particular bacteriochlorin, corresponding to formula (II), is the absorption of light in the red region at 750 nm, as shown in figure 2. At this wavelength, the effects of screening caused by human tissues, in particular hemoglobin, are not so pronounced, which makes it possible to increase the "amount" of light reaching the photosensitizer inside the cells.

The present invention also relates to derivatives corresponding to the formula (II), where

- X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , each, identical or different, are selected from halogen atoms and / or hydrogen atoms so that at least one of X radicals in each pair of X ¹ / X ² , X ³ / X ⁴ , X ⁵ / X ⁶ , X ⁷ / X ⁸ is a halogen atom.

As in the case of chlorin described above, the presence of halogen atoms in the ortho positions of the phenyl substituent increases the quantum yield of the formation of the triplet state without significantly reducing its half-life and, therefore, also increases the formation of singlet oxygen.

The present invention also relates to certain derivatives corresponding to the formula (II), where

- X ² , X ⁴ , X ⁶ and X ⁸ are either chlorine atoms or fluorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are chlorine atoms.

According to another embodiment of the present invention and in accordance with formula (II):

- X ² , X ⁴ , X ⁶ and X ⁸ are either chlorine atoms or fluorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are OH groups.

In addition, the present invention relates to derivatives corresponding to the formula (II), where X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms, R ¹ , R ² , R ³ , R ⁴ are either chlorine atoms or OH groups.

The presence of halogen atoms in the ortho positions of the phenyl substituent also makes tetrapyrrole macrocycles and their derivatives more stable.

According to the present invention, halogen atoms are fluorine and / or chlorine and / or bromine.

Another objective of the present invention is a method for producing the derivatives described above, comprising the following steps:

- the stage of chlorosulfonation of the corresponding halogenated porphyrin,

- the stage of recovery of halogenated and chlorosulfonated porphyrin to chlorin and / or to bacteriochlorin using hydrazide in the presence of inhibited organic bases,

- the stage of interaction between the chlorosulfonated part of the molecule and amines and / or amino acids and / or alcohols or the stage of hydrolysis of chlorine of the chlorosulfonyl residue with water.

Using a non-limiting example, chemical synthesis of a derivative corresponding to formula (I), where

- X ² , X ⁴ , X ⁶ and X ⁸ are chlorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are OH groups,

carried out as described below:

5,10,15,20-tetrakis (2-chloro-3-chlorosulfonylphenyl) porphyrin (0.0658 g, 6.55 × 10 ^-5 mol) was previously dissolved in N, N-dimethylformamide and / or toluene and / or pyridine and / or picoline (50 ml), previously dehydrated and distilled, the temperature of the reaction mixture was raised from 50 ° C to 150 ° C. Then, a solution of p-toluene-sulfonyl hydrazide (5.5 × 10 ^-5 mol) and a bulk organic base such as DABCO and / or DBU in DMF (5 ml) were added to the reaction mixture. The mixture was stirred at 150 ° C. under nitrogen and in the absence of light.

The progress of the reaction was monitored using the spectrophotometric method in the visible region until the Q band at 650 nm reached a maximum.

Then the solvent was evaporated, the reaction mixture was treated, the resulting product was hydrolyzed with water and purified by extraction from solution into solution. Recrystallization with methanol / dichloromethane gave the product in 80% yield. Mass Spectroscopy (FAB), m / z: 1075 (central peak).

Visible ultraviolet region of the spectrum (buffer solution, NaOH, KH ₂ P0 ₄ ), maximum wavelength (in nm): 414, 516, 542, 598, 649. Fluorescence quantum yield: 0.04.

Half-life of the triplet state in a nitrogen-saturated aqueous solution: 235 μs.

Half-life of the triplet state in an air-saturated aqueous solution: 3.88 μs.

Quantum yield of singlet oxygen formation in a deuterated aqueous solution: 0.56.

Half-life of singlet oxygen: 65 μs in deuterated water.

Schematically, the reaction can be represented as follows:

Also, using a non-limiting example, the chemical synthesis of a derivative corresponding to formula (II), where

- X ² , X ⁴ , X ⁶ and X ⁸ are chlorine atoms;

- X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,

- R ¹ , R ² , R ³ , R ⁴ are OH groups,

carried out as described below:

5,10,15,20-tetrakis (2-chloro-3-chlorosulfonylphenyl) porphyrin (0.0658 g, 6.55 × 10 ^-5 mol) was previously dissolved in N, N-dimethylformamide and / or in toluene and / or in pyridine and / or picoline (50 ml), previously dehydrated and distilled, the temperature of the reaction mixture was raised from 50 ° C to 150 ° C. Then a solution of p-toluene-sulfonyl hydrazide (130.0 × 10 ^-5 mol) and DABCO in DMF (5 ml) was added to the reaction mixture. The mixture was stirred at 150 ° C. under nitrogen and protected from light. The progress of the reaction was monitored using a spectrophotometric method in the visible region until the Q band at 750 nm reached a maximum.

Then the solvent was evaporated, the reaction mixture was treated, the resulting product was hydrolyzed, and the desired product was obtained with a yield of 77%. Mass Spectroscopy (FAB), m / z: 1077 (central peak).

Visible ultraviolet region of the spectrum (buffer solution, NaOH, KH ₂ P0 ₄ ), maximum wavelength (in nm): 353, 376, 486, 518, 604, 748.

Fluorescence quantum yield: 0.013.

Half-life of the triplet state in a nitrogen-saturated aqueous solution: 235 μs.

Half-life of the triplet state in an air-saturated aqueous solution: 3.88 μs.

Quantum yield of singlet oxygen formation in a deuterated aqueous solution: more than 30%.

Half-life of singlet oxygen: 65 μs in deuterated water.

Also, schematically, the reaction can be represented as follows:

5,10,15,20-tetrakis (2-chloro-3-chlorosulfonylphenyl) porphyrin, used as the starting porphyrin in the synthesis of chlorin and bacteriochlorin described above, is known to those skilled in the art and is described in the literature, in particular in the Journal of Heterocyclic Chemistry, 28: 635 (1991).

According to the present invention, the quantum yields of singlet oxygen formation were obtained by comparing the intensity of singlet oxygen phosphorescence at a wavelength of 1270 nm in a solution containing the photosensitizer under study and the phosphorescence intensity obtained from another solution having the same absorption at the same wavelength and containing the compared photosensitizer.

According to the present invention, a meso-tetrakis (2-chloro-3-sulfophenyl) porphyrin (TPPS) photosensitizer was used for comparison, whose quantum yield of singlet oxygen formation in deuterated water was 0.64 at pH-7.4 (Photochem. Photobiol., 70: 391, 1999).

The present invention also relates to anticancer and / or antiviral and / or antimicrobial drugs used to treat humans or animals, containing, as the main active substance, one or more compounds described in the present invention.

This type of drug, used in particular in photodynamic therapy, may also contain one or more pharmacologically acceptable excipients.

Of course, other embodiments of the present invention available to a person skilled in the art can be considered without departing from the basis of the invention.

Claims (19)

1. A derivative of porphyrin, in particular chlorin, having the formula (I)

where X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ each, identical or different, are selected from halogen atoms and / or hydrogen atoms, so that at least one of X radicals in each pair of X ¹ / X ² , X ³ / X ⁴ , X ⁵ / X ⁶ , X ⁷ / X ⁸ is a halogen atom,
R ¹ , R ² , R ³ , R ⁴ each, identical or different, are selected from the following substituents: OH groups, amino acids, OR groups and NHR groups and / or a chlorine atom, where R is an alkyl group having from 1 to 12 carbon atoms. 2. The porphyrin derivative according to claim 1, corresponding to the formula (I), where
X ^2, X ^4, X ⁶ and X ⁸ are chlorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are chlorine atoms. 3. The porphyrin derivative according to claim 1, corresponding to the formula (I), where
X ² , X ⁴ , X ⁶ and X ⁸ are fluorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ^1, R ^2, R ^3, R ⁴ are chlorine atoms. 4. The porphyrin derivative according to claim 1, corresponding to the formula (I), where
X ² , X ⁴ , X ⁶ and X ⁸ are fluorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are OH groups. 5. The porphyrin derivative according to claim 1, corresponding to the formula (I), where
X ² , X ⁴ , X ⁶ and X ⁸ are fluorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are OH groups. 6. The porphyrin derivative according to claim 1, corresponding to the formula (I), where
X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms;
R ¹ , R ² , R ³ , R ⁴ are chlorine atoms. 7. The derivative of porphyrin according to claim 1, corresponding to the formula (I), where
X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms;
R ¹ R ² , R ³ , R ⁴ are OH groups. 8. A derivative of porphyrin, in particular bacteriochlorin, having the formula (II)

where X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ each, identical or different, are selected from halogen atoms and / or hydrogen atoms;
R ¹ , R ² , R ³ , R ⁴ each, identical or different, are selected from the following substituents: OH groups, amino acids, OR groups and NHR groups and / or chlorine atoms, where R is an alkyl group having from 1 to 12 carbon atoms. 9. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ each, identical or different, are selected from halogen atoms and / or hydrogen atoms, so that at least one of the radicals X in each pair X ¹ / X ² , X ³ / X ⁴ , X ⁵ / X ⁶ , X ⁷ / X ⁸ is a halogen atom. 10. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ^2, X ^4, X ⁶ and X ⁸ are chlorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are chlorine atoms. 11. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ² , X ⁴ , X ⁶ and X ⁸ are fluorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are chlorine atoms. 12. The porphyrin derivative of claim 8, corresponding to the formula (II), where
X ² , X ⁴ , X ⁶ and X ⁸ are chlorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are OH groups. 13. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ² , X ⁴ , X ⁶ and X ⁸ are fluorine atoms;
X ¹ , X ³ , X ⁵ and X ⁷ are hydrogen atoms,
R ¹ , R ² , R ³ , R ⁴ are OH groups. 14. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms;
R ¹ , R ² , R ³ , R ⁴ are chlorine atoms. 15. The derivative of porphyrin according to claim 8, corresponding to the formula (II), where
X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ are chlorine atoms;
R ¹ , R ² , R ³ , R ⁴ are OH groups. 16. The porphyrin derivative according to any one of claims 1 or 8, characterized in that the halogen atoms are fluorine and / or chlorine and / or bromine. 17. A method of obtaining a porphyrin derivative according to any one of the preceding paragraphs, characterized in that it includes:
the step of reducing the corresponding halogenated and chlorosulfonated porphyrin to chlorin and / or bacteriochlorin using hydrazide and a bulky organic base such as DABCO (1,4-diazabicyclo [2.2.2] octane) and / or DBU (1,8-diazabicyclo [5.4. 0] undec-7-en);
the stage of interaction between the chlorosulfonyl group and amines and / or amino acids and / or alcohols. 18. A method of obtaining a derivative according to any one of the preceding paragraphs, characterized in that it includes:
a step of reducing the corresponding halogenated and chlorosulfonated porphyrin to chlorin and / or bacteriochlorin using hydrazide and a bulky organic base such as DABCO and / or DBU;
the stage of hydrolysis of chlorine of the chlorosulfonyl group. 19. An anti-cancer drug for use in humans or animals, containing as the main active substance one or more compounds according to claims 1-16.

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